Synaptopodin antibody

Manufactured by Santa Cruz Biotechnology

Synaptopodin antibody is a protein-specific antibody used in research applications to detect and analyze the presence and distribution of synaptopodin, a cytoskeletal protein found in podocytes and dendritic spines. The antibody can be utilized in techniques such as immunohistochemistry, immunocytochemistry, and Western blotting to support research in cell biology, neuroscience, and nephrology.

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Lab products found in correlation

Complete ultra, by Roche (2 mentions) Secondary antibody conjugated with hrp, by Thermo Fisher Scientific (2 mentions) Gapdh antibody, by Santa Cruz Biotechnology (2 mentions) Micro bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Nitrocellulose membrane, by Bio-Rad (2 mentions) Westernbright ecl, by Advansta (2 mentions) Monoclonal antibodies to the bkv major capsid protein vp1, by Santa Cruz Biotechnology (1 mentions) Nephrin antibody, by Abcam (1 mentions) Myc tag antibody, by Cell Signaling Technology (1 mentions) Protease phosphatase inhibitor cocktail, by Cell Signaling Technology (1 mentions) Mouse monoclonal β actin antibody, by Merck Group (1 mentions) Mouse monoclonal wt1 antibody, by Santa Cruz Biotechnology (1 mentions) Phenyl methyl sulfonyl fluoride (pmsf), by Merck Group (1 mentions) Lysis buffer, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

Synaptopodin (28 citations) Anti-synaptopodin antibody (2 citations) Anti-synaptopodin primary antibody (2 citations) Goat anti-synaptopodin antibody (2 citations)

3 protocols using synaptopodin antibody

Western Blot Analysis of Polyoma Virus Protein

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Cell extracts were prepared using RIPA lysis buffer [50 mM Tris pH 7.5, 150 mM NaCl, 2 mM ethylenediaminetetraacetic acid (EDTA) pH 8.0, 1% NP40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS), and proteinase inhibitor (cOmplete ULTRA, Roche, Basel, Switzerland). Lysates were incubated on ice for 30 minutes and then clarified by centrifugation. Total protein was measured using a micro BCA protein assay kit (ThermoFisher Scientific). Protein lysates (30 μg) were separated by 10% SDS-PAGE, transferred to nitrocellulose membranes (Bio-Rad), blocked with 5% milk in 0.1% TBST (0.1% Tween 20, 20 mM Tris, 150 mM NaCl), and incubated at 4 °C overnight with monoclonal antibodies to the BKV major capsid protein VP1 (Santa Cruz Biotech) at 1:250 dilution. A synaptopodin antibody (Santa Cruz Biotech) was used at 1:250 dilution and the GAPDH antibody (Santa Cruz Biotech) was used at 1:3000 dilution. Membranes were washed five times in 0.1% TBST and incubated for one hour with a corresponding secondary antibody conjugated with HRP (ThermoFisher Scientific) at a dilution of 1:50,000. Immunoreactive bands were detected with WesternBright ECL (Advansta, San Jose, CA) following exposure to X-ray film.

Popik W., Khatua A.K., Fabre N.F., Hildreth J.E, & Alcendor D.J. (2019). BK Virus Replication in the Glomerular Vascular Unit: Implications for BK Virus Associated Nephropathy. Viruses, 11(7), 583.

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Western Blot Analysis of Flavivirus Proteins

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Cell extracts were prepared using RIPA lysis buffer [50 mM Tris pH 7.5, 150 mM NaCl, 2 mM ethylenediaminetetraacetic acid (EDTA) pH 8.0, 1% NP40, 0.5% sodium deoxycholate, 0.1% sodium dodecyl sulfate (SDS), and proteinase inhibitor (Complete Ultra, Roche). Lysates were incubated on ice for 30 min and then clarified by centrifugation. Total protein was measured by micro BCA protein assay kit (ThermoFisher Scientific). Protein lysates (30 μg) were separated by 10% SDS-PAGE, transferred to nitrocellulose membranes (Bio-Rad), blocked with 5% milk in 0.1% TBST (0.1% Tween 20, 20 mM Tris, 150 mM NaCl) and incubated at 4 °C overnight with 4 G-2 Flavivirus group antigen monoclonal antibody (Millipore, Temecula, CA, USA) at 1:250 dilution. Synaptopodin antibody (Santa Cruz Biotechnology) was used at 1:250 dilution and GAPDH antibody (Santa Cruz Biotechnology) at 1:3000 dilution. Membranes were washed five times in 0.1% TBST and incubated for one hour with corresponding secondary antibody conjugated with HRP (ThermoFisher Scientific) at a dilution of 1:50,000. Immunoreactive bands were detected with WesternBright ECL (Advansta) following exposure to X-ray film.

Popik W., Khatua A., Hildreth J.E., Lee B, & Alcendor D.J. (2018). Phosphorodiamidate morpholino targeting the 5′ untranslated region of the ZIKV RNA inhibits virus replication. Virology, 519, 77-85.

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Podocyte Protein Expression Analysis

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Conditionally immortalized podocyte were differentiated per established protocols (PMID: 19955187), harvested, and treated with lysis buffer (Pierce Biotechnology, Rockford, IL) supplemented with phosphatase/protease inhibitor cocktail, 1:100 dilution (Cell Signaling Technology) and 1:100 PMSF (Sigma-Aldrich) for 15 min on ice. Cells were then spun at 14,000 RPM for 10 mins at 4 °C (Eppendorf). Protein Immunoblotting was performed as described previously using a MYC tag antibody 1:000 (Cell Signaling Technology), rabbit polyclonal nephrin antibody1:1000 (Abcam), mouse monoclonal Glepp1 antibody 1:1000 (Santa Cruz), mouse monoclonal WT1 antibody 1:300 (Santa Cruz), rabbit polyclonal CD2AP 1:1000 (Abcam), rabbit polyclonal Synaptopodin antibody 1:800 (Santa Cruz), rabbit polyclonal INF2 antibody 1:1000 (Bethyl Laboratories), and mouse monoclonal β-actin antibody 1:3000 (Sigma-Aldrich)10 (link).

Hall G., Lane B., Chryst-Ladd M., Wu G., Lin J.J., Qin X., Hauser E.R, & Gbadegesin R. (2017). Dysregulation of WTI (−KTS) is Associated with the Kidney-Specific Effects of the LMX1B R246Q Mutation. Scientific Reports, 7, 39933.

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